BRENDA - Enzyme Database show
show all sequences of 1.1.1.169

Crystal structure of Escherichia coli ketopantoate reductase at 1.7 A resolution and insight into the enzyme mechanism

Matak-Vinkovic, D.; Vinkovic, M.; Saldanha, S.A.; Ashurst, J.L.; von Delft, F.; Inoue, T.; Miguel, R.N.; Smith, A.G.; Blundell, T.L.; Abell, C.; Biochemistry 40, 14493-14500 (2001)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli BL21(DE3)
Escherichia coli
Crystallization (Commentary)
Crystallization
Organism
prismic crystals, hanging drop vapor-diffusion technique
Escherichia coli
Engineering
Amino acid exchange
Commentary
Organism
additional information
Lys176 and Glu256 important for binding ketopantoate and the catalytic mechanism
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.004
-
NADPH
-
Escherichia coli
0.02
-
NADPH
-
Escherichia coli
0.06
-
ketopantoate
-
Escherichia coli
0.12
-
ketopantoate
-
Escherichia coli
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
does not require Mg2+, independent of divalent metal ions
Escherichia coli
additional information
-
Stenotrophomonas maltophilia
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
30500
-
3-5 * 30500 or 30000, SDS-PAGE
Stenotrophomonas maltophilia
31100
-
1 * 31100, HPLC gel filtration
Salmonella enterica subsp. enterica serovar Typhimurium
33000
-
SDS-PAGE
Escherichia coli
34000
-
1 * 34000
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ketopantoate + NADPH
Salmonella enterica subsp. enterica serovar Typhimurium
-
pantoate + NADP+
-
Salmonella enterica subsp. enterica serovar Typhimurium
?
ketopantoate + NADPH
Escherichia coli
pantothenate biosynthetic pathway
pantoate + NADP+
-
Escherichia coli
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
P0A9J4
expressed in BL21(DE3)
-
Salmonella enterica subsp. enterica serovar Typhimurium
-
-
-
Stenotrophomonas maltophilia
-
845
-
Stenotrophomonas maltophilia 845
-
845
-
Posttranslational Modification
Posttranslational Modification
Commentary
Organism
side-chain modification
SeMet substitution
Escherichia coli
Purification (Commentary)
Commentary
Organism
FPLC, anion exchange chromatography, gel filtration
Escherichia coli
Specific Activity [micromol/min/mg]
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
additional information
-
-
Escherichia coli
additional information
-
3.8-fold higher specific activity with NADPH than with NADH
Salmonella enterica subsp. enterica serovar Typhimurium
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ketopantoate + NADPH
-
286110
Salmonella enterica subsp. enterica serovar Typhimurium
pantoate + NADP+
-
286110
Salmonella enterica subsp. enterica serovar Typhimurium
?
ketopantoate + NADPH
pantothenate biosynthetic pathway
286110
Escherichia coli
pantoate + NADP+
-
286110
Escherichia coli
?
additional information
-
286110
Escherichia coli
?
-
-
-
-
additional information
3.8-fold higher specific activity with NADPH than with NADH
286110
Salmonella enterica subsp. enterica serovar Typhimurium
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
monomer
1 * 33000, gel filtration; 1 * 34000
Escherichia coli
monomer
1 * 31100, HPLC gel filtration
Salmonella enterica subsp. enterica serovar Typhimurium
oligomer
3-5 * 30500 or 30000, SDS-PAGE
Stenotrophomonas maltophilia
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
8
-
ketopantoate
-
Escherichia coli
40
-
ketopantoate
-
Escherichia coli
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
-
Salmonella enterica subsp. enterica serovar Typhimurium
NADPH
-
Stenotrophomonas maltophilia
NADPH
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli BL21(DE3)
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
-
Salmonella enterica subsp. enterica serovar Typhimurium
NADPH
-
Stenotrophomonas maltophilia
NADPH
-
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
Organism
prismic crystals, hanging drop vapor-diffusion technique
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
Lys176 and Glu256 important for binding ketopantoate and the catalytic mechanism
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.004
-
NADPH
-
Escherichia coli
0.02
-
NADPH
-
Escherichia coli
0.06
-
ketopantoate
-
Escherichia coli
0.12
-
ketopantoate
-
Escherichia coli
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
does not require Mg2+, independent of divalent metal ions
Escherichia coli
additional information
-
Stenotrophomonas maltophilia
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
30500
-
3-5 * 30500 or 30000, SDS-PAGE
Stenotrophomonas maltophilia
31100
-
1 * 31100, HPLC gel filtration
Salmonella enterica subsp. enterica serovar Typhimurium
33000
-
SDS-PAGE
Escherichia coli
34000
-
1 * 34000
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ketopantoate + NADPH
Salmonella enterica subsp. enterica serovar Typhimurium
-
pantoate + NADP+
-
Salmonella enterica subsp. enterica serovar Typhimurium
?
ketopantoate + NADPH
Escherichia coli
pantothenate biosynthetic pathway
pantoate + NADP+
-
Escherichia coli
?
Posttranslational Modification (protein specific)
Posttranslational Modification
Commentary
Organism
side-chain modification
SeMet substitution
Escherichia coli
Purification (Commentary) (protein specific)
Commentary
Organism
FPLC, anion exchange chromatography, gel filtration
Escherichia coli
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
additional information
-
-
Escherichia coli
additional information
-
3.8-fold higher specific activity with NADPH than with NADH
Salmonella enterica subsp. enterica serovar Typhimurium
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ketopantoate + NADPH
-
286110
Salmonella enterica subsp. enterica serovar Typhimurium
pantoate + NADP+
-
286110
Salmonella enterica subsp. enterica serovar Typhimurium
?
ketopantoate + NADPH
pantothenate biosynthetic pathway
286110
Escherichia coli
pantoate + NADP+
-
286110
Escherichia coli
?
additional information
-
286110
Escherichia coli
?
-
-
-
-
additional information
3.8-fold higher specific activity with NADPH than with NADH
286110
Salmonella enterica subsp. enterica serovar Typhimurium
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 33000, gel filtration; 1 * 34000
Escherichia coli
monomer
1 * 31100, HPLC gel filtration
Salmonella enterica subsp. enterica serovar Typhimurium
oligomer
3-5 * 30500 or 30000, SDS-PAGE
Stenotrophomonas maltophilia
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
8
-
ketopantoate
-
Escherichia coli
40
-
ketopantoate
-
Escherichia coli
Other publictions for EC 1.1.1.169
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [░C]
Temperature Range [░C]
Temperature Stability [░C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [░C] (protein specific)
Temperature Range [░C] (protein specific)
Temperature Stability [░C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
725293
Miller
PanG, a new ketopantoate reduc ...
Francisella tularensis subsp. novicida
J. Bacteriol.
195
965-976
2013
-
-
-
-
-
-
-
-
-
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
736827
Tomita
Identification and characteriz ...
Thermococcus kodakarensis, Thermococcus kodakarensis KUW1
Mol. Microbiol.
90
307-321
2013
-
-
1
-
1
-
1
7
-
-
2
4
-
8
-
-
1
-
-
-
-
-
13
1
1
1
1
6
1
-
-
4
1
-
-
-
-
1
4
-
1
-
-
1
1
7
-
-
2
4
-
-
-
1
-
-
-
-
13
1
1
1
1
6
1
-
-
-
-
2
2
-
6
6
696603
Headey
Backbone assignments of the 34 ...
Escherichia coli
Biomol. NMR Assign.
2
93-96
2008
-
-
1
-
-
-
-
-
-
-
1
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
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-
-
-
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1
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
684140
Ciulli
pH-tuneable binding of 2-phosp ...
Escherichia coli
Acta Crystallogr. Sect. D
63
171-178
2007
-
-
-
1
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
687675
Ciulli
Crystal structure of Escherich ...
Escherichia coli
J. Biol. Chem.
282
8487-8497
2007
-
-
1
1
6
-
-
3
-
-
-
1
-
2
-
-
-
-
-
-
-
-
2
-
1
-
-
2
1
-
-
2
-
-
-
-
-
1
2
1
6
-
-
-
-
3
-
-
-
1
-
-
-
-
-
-
-
-
2
-
1
-
-
2
1
-
-
-
-
-
-
-
-
-
669806
Ciulli
Probing hot spots at protein-l ...
Escherichia coli
J. Med. Chem.
49
4992-5000
2006
-
-
-
-
-
-
7
1
-
-
-
1
-
2
-
-
-
1
-
-
-
-
2
-
-
-
-
-
1
-
-
3
6
-
-
-
-
-
3
-
-
-
-
7
6
1
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
667561
Ciulli
Biophysical tools to monitor e ...
Escherichia coli
Biochem. Soc. Trans.
33
767-771
2005
-
-
-
-
-
-
1
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
667687
Lobley
The crystal structure of Esche ...
Escherichia coli
Biochemistry
44
8930-8939
2005
-
-
1
1
7
-
-
1
-
-
-
1
-
2
-
-
1
1
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
1
7
-
-
-
-
1
-
-
-
1
-
-
-
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
654677
Zheng
Substrate specificity and kine ...
Escherichia coli
Biochemistry
42
11289-11296
2003
-
-
1
-
-
-
-
8
-
-
-
1
-
2
-
-
-
-
-
-
-
-
7
-
1
-
-
-
1
1
-
4
-
-
-
-
-
1
4
-
-
-
-
-
-
8
-
-
-
1
-
-
-
-
-
-
-
-
7
-
1
-
-
-
1
1
-
-
-
-
-
-
-
-
656330
Merkamm
Ketopantoate reductase activit ...
Corynebacterium glutamicum
J. Biotechnol.
104
253-260
2003
-
-
1
-
-
-
-
1
-
-
-
1
-
3
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286110
Matak-Vinkovic
Crystal structure of Escherich ...
Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium, Stenotrophomonas maltophilia, Stenotrophomonas maltophilia 845
Biochemistry
40
14493-14500
2001
-
-
1
1
1
-
-
4
-
2
4
2
-
7
-
1
1
-
-
-
2
-
4
3
-
-
-
2
-
-
-
3
-
-
-
-
-
1
3
1
1
-
-
-
-
4
-
2
4
2
-
-
1
1
-
-
2
-
4
3
-
-
-
2
-
-
-
-
-
-
-
-
-
-
286108
Zheng
Kinetic and mechanistic analys ...
Escherichia coli
Biochemistry
39
3708-3717
2000
-
-
1
-
-
-
2
4
-
1
4
1
-
2
-
-
1
-
-
-
1
-
2
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
-
-
-
2
-
4
-
1
4
1
-
-
-
1
-
-
1
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286109
Zheng
Identification of active site ...
Escherichia coli
Biochemistry
39
16244-16251
2000
5
-
1
-
10
-
-
12
-
1
1
-
-
2
-
-
1
-
-
-
1
-
2
1
-
-
-
-
1
-
-
2
-
-
-
5
-
1
2
-
10
-
-
-
-
12
-
1
1
-
-
-
-
1
-
-
1
-
2
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
286106
Elischewski
Pantothenate production in Esc ...
Escherichia coli
J. Biotechnol.
75
135-146
1999
-
-
1
-
-
-
-
-
-
-
1
-
-
3
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286107
Frodyma
ApbA, the ketopantoate reducta ...
Escherichia coli, Escherichia coli BL21/lambdaDE3, Saccharomyces cerevisiae, Salmonella enterica subsp. enterica serovar Typhimurium, Stenotrophomonas maltophilia 845, Stenotrophomonas maltophilia
J. Biol. Chem.
273
5572-5576
1998
-
-
1
-
-
-
-
2
-
-
1
3
-
10
-
-
1
-
-
-
6
-
12
2
1
-
-
-
1
-
-
8
-
-
-
-
-
1
8
-
-
-
-
-
-
2
-
-
1
3
-
-
-
1
-
-
6
-
12
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
286102
Kataoka
-
Novel enzymic production of D- ...
Agrobacterium sp., Agrobacterium tumefaciens, Pseudomonas putida, Stenotrophomonas maltophilia
Agric. Biol. Chem.
54
177-182
1990
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
3
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
286103
Shimizu
Ketopantoic acid reductase of ...
Saccharomyces cerevisiae, Stenotrophomonas maltophilia, Stenotrophomonas maltophilia 845
J. Biol. Chem.
263
12077-12084
1988
-
-
-
1
-
-
10
4
-
1
4
-
-
6
-
-
1
-
-
-
2
1
6
1
1
1
3
-
2
-
2
4
-
-
-
-
-
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4
1
-
-
-
10
-
4
-
1
4
-
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-
-
1
-
-
2
1
6
1
1
1
3
-
2
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2
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286105
Wilken
Ketopantoic acid and ketopanto ...
Escherichia coli, Saccharomyces cerevisiae
J. Biol. Chem.
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2311-2314
1975
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2
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2
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2
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2
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2
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2
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2
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286104
King
-
Separation and preliminary stu ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae NRRL Y-2034
J. Biol. Chem.
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4096-4098
1972
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2
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1
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4
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2
1
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2
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2
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1
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2
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4
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2
1
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